For lipid quantification by shotgun MS, 700 μL of a mixture of internal standards in MTBE/MeOH (5:1.5; [vol/vol]) were added to the dried lipid fractions. MS analyses were performed as previously described (74 (link)) on a Q Exactive instrument (Thermo Fisher Scientific) equipped with a robotic nanoflow ion source TriVersa NanoMate (Advion BioSciences) using nanoelectrospray chips with the diameter of spraying nozzles of 4.1 μm. The ion source was controlled by the Chipsoft 8.3.1 software (Advion BioSciences). Spectra was filtered based on repetition rate as previously described and analyzed by a laboratory-developed script (75 (link), 76 (link)). Lipids were identified by LipidXplorer software (77 (link)).
Bedard M., Shrestha D., Priestman D.A., Wang Y., Schneider F., Matute J.D., Iyer S.S., Gileadi U., Prota G., Kandasamy M., Veerapen N., Besra G., Fritzsche M., Zeissig S., Shevchenko A., Christianson J.C., Platt F.M., Eggeling C., Blumberg R.S., Salio M, & Cerundolo V. (2019). Sterile activation of invariant natural killer T cells by ER-stressed antigen-presenting cells. Proceedings of the National Academy of Sciences of the United States of America, 116(47), 23671-23681.
Other organizations :
MRC Human Immunology Unit, Medical Research Council, University of Oxford, TU Dresden, Max Planck Institute of Molecular Cell Biology and Genetics, Brigham and Women's Hospital, Harvard University, Massachusetts General Hospital, University of Birmingham, Nuffield Orthopaedic Centre, Fraunhofer Institute for Applied Optics and Precision Engineering, Leibniz Institute of Photonic Technology
Mixture of internal standards in MTBE/MeOH (5:1.5; [vol/vol])
Q Exactive instrument (Thermo Fisher Scientific) equipped with a robotic nanoflow ion source TriVersa NanoMate (Advion BioSciences) using nanoelectrospray chips with the diameter of spraying nozzles of 4.1 μm
Ion source controlled by the Chipsoft 8.3.1 software (Advion BioSciences)
Spectra filtered based on repetition rate as previously described
Lipids identified by LipidXplorer software
controls
No positive or negative controls were explicitly mentioned in the provided information.
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